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We narrowed to 79 results for: plasmid cloning by pcr

Showing: 61 - 70 of 79 results

  1. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    Published
    April 19, 2017, 3:25 p.m.
    ...Joe Mellor, Founder and CEO of seqWell Inc. Plasmids and PCR products are the bread and butter of molecular... about the presence of non-plasmid DNA or heterogeneous and/or mixed clones present in the original sample... circularized plasmids from plexWell™ NGS data. Once Addgene produces plates of plasmid DNA, they are ...using these methods for routine processes like plasmid validation. At seqWell, Inc., our mission is to...Addgene’s large and growing collection of curated plasmids from all over the world. The rest of this piece...partnership with Addgene to sequence large numbers of plasmids. plexWell™ technology overview For any NGS experiment...individual input DNA samples (e.g. a collection of 96 plasmids on a plate) will typically have a wide range of...

  2. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    Published
    Dec. 7, 2020, 2:53 p.m.
    ... involving two-plasmid or single-plasmid systems (Vo et al., 2020). In the two-plasmid system, an effector... follows a simple workflow: Clone spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon...Transform plasmid(s) into cells and plate overnight Screen colonies phenotypically or by PCR/sequencing...group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express the...effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under the same promoter...promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and ...

  3. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    Published
    July 7, 2015, 3:36 p.m.
    ...remove the selection marker. Other labs have used PCR-based screening to detect mutants, but this method... To use this SEC-based system, first design and clone 500-700 bp homology arms for your gene of interest...This construct will serve as the repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins...proteins. The repair template and Cas9-sgRNA plasmid is then injected into the germline of young adult ....115.178335. [Epub ahead of print] PubMed. Find plasmids from this publication at Addgene. Other strategies...10.1038/nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at Addgene. Scalable and...genetics.114.170423. Epub 2014 Sep 23. PubMed. Find plasmids from this publication at Addgene. Additional...

  4. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    Published
    July 21, 2020, 1:15 p.m.
    ...to elute and store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on...Reference Find plasmids for your research Browse protocols for molecular biology, cloning, and more ...This method is best used for DNA fragments (like a PCR product). This method also gives an indicator of ...studies and is not used as much for the standard plasmid prep. Diphenylamine method Another absorbance based...

  5. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...using a gRNA, which can easily be cloned into a gRNA expression plasmid. Experimental use Since the initial... tag expressed on a plasmid, allowing for in vivo biotinylation through a plasmid-based biotin ligase....CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...bound genomic locus, which they then confirmed by PCR. Subsequent mass spectroscopy studies allowed for...specific genomic locus, which can require multiple cloning steps (Fujita et al., 2013). CRISPR-mediated purification...currently no cure (Campbell et al., 2018). Find plasmids from the Fujii lab! Conclusions CRISPR-mediated...

  6. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    Published
    June 23, 2016, 2:30 p.m.
    ...expression plasmid encoding the ORF to be expressed Finally, the researcher needs a plasmid encoding their...PhosTag system (see below). This plasmid backbone can also be cloned with an ORF of interest for customized...E17TAG GFP plasmid - Addgene #68295 SupD plasmid – Addgene # 68307 For further technical reference and protocols...Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein Purification...protocols Sep-OTSλ plasmid - Addgene # 68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG...strain for protein production, and an expression plasmid encoding the open reading frame (ORF) of interest...translation of pSer-incorporated proteins on a single plasmid. This includes 4 copies of a tRNA that specifically...

  7. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...blog Addgene's Tips for Plasmid Quality Control Plasmids 101: In-house NGS Plasmid Quality Control Now at...to share. However, the cloning process — including the propagation of plasmids in bacteria — has many ...sequencing all incoming plasmids. If you are interested in requesting one of these plasmids, you can request ...data for each of their plasmids, from descriptions of key components of the plasmids to partial, full, or...to ensure all plasmids pass our high-quality standards and contain the expected plasmid features.  ...Once Addgene receives deposited plasmids, they undergo complete plasmid sequencing using an in-house Illumina...will compare the NGS results to the plasmid information, other plasmids in the deposit, NCBI sequence and...

  8. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    .... citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering...consists of two plasmids: pCas9: carries Cas9 and chloramphenicol resistance pCRISPR: carries a spacer...other advantages including shorter plasmid curing time. pCRISPomyces for editing in Streptomyces  Streptomyces...engineering functions in bacteria, but have no fear - plasmids from Addgene depositors are making it easier than...and these rare events are easily detectable using PCR. The system also functions in S. pneumoniae and can...gRNA and a backbone-targeting gRNA cassette for plasmid curing through "self-destruction" after induction...editing takes two days, and the pTargetF and pCas plasmids can be subsequently cured from the bacteria. Although...

  9. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...insertions/deletions using a mismatch repair assay? Or is PCR followed by gel electrophoresis and/or next-generation...add a tag, you will also need to deliver a donor plasmid or single-stranded DNA oligonucleotide, so you ...generate stable cell lines. Packaging and Envelope plasmids provide the necessary components to make lentiviral...for in vivo viral delivery. Cas9 mRNA and gRNA Plasmids containing gRNA and Cas9 are used in in vitro ...efficiency of CRISPR delivery. There are a few plasmids at Addgene that have been published as CRISPR ...evaluate component delivery and sgRNA efficacy by cloning in genome target sequence into EGFP reporter. Target...User) Resources on Addgene.org Browse CRISPR Plasmids for Mammalian Systems Check out our CRISPR Guide...

  10. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...all CRISPR plasmids and resources at Addgene Find CRISPR base editors Find CRISPR plasmids for plants ... al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA (protospacer+scaffold) in a...single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables us to target...restriction enzyme (RE) recognition site, RE analysis of PCR product could be used to detect success. Although...
Showing: 61 - 70 of 79 results